gut microbiotainvolved mechanisms in enhancing - exposure ... · gut microbiotainvolved mechanisms...
TRANSCRIPT
Gut microbiota-involved mechanisms in enhancing systemic exposure of ginsenosides by coexisting polysaccharides in
ginseng decoction
Shan-Shan Zhou1, 2, #, Jun Xu3, #, He Zhu1, Jie Wu2, Jin-Di Xu1, Ru Yan4, Xiu-Yang Li1, 2, Huan-Huan Liu1, 2, Su-Min Duan2, Zhuo Wang2, Hu-Biao Chen3, Hong Shen2, *,
Song-Lin Li1, 2, *
Supplementary Table 1 Linearity, sensitivity and contents of the eleven ginsenosides in DST (n=3)
Analyte Linear range (ng/mL) Equation R2 LOD (ng/mL) LOQ (ng/mL) Contents (μg/mL)
Re 1.00-200 y=5.23x+29.63 0.9990 0.50 1.00 225.70 ± 5.10
Rg1 1.00-200 y=7.15x+239.04 0.9991 0.50 1.00 360.69 ± 17.53
Rf 1.00-200 y=19.28x+51.01 0.9996 0.50 1.00 107.72 ± 2.35
Rb1 1.00-200 y=942.97x+5804.09 0.9990 0.50 1.00 52.01 ± 1.18
20(S)-Rg2 1.00-200 y=2.06x+7.10 0.9992 0.50 1.00 83.83 ± 2.58
Rc 1.00-200 y=33.13x+503.00 0.9990 0.50 1.00 190.51 ± 9.18
Rb2 1.00-200 y=42.02x+537.50 0.9991 0.50 1.00 198.54 ± 7.87
Rd 1.00-200 y=2.84x+1.95 0.9992 0.50 1.00 72.36 ± 2.82
F2 1.00-200 y=20.49x+31.73 0.9998 0.50 1.00 1.08 ± 0.13
20(S)-Rg3 5.00-200 y=10.12x+11.24 0.9999 2.50 5.00 138.48 ± 6.85
CK 1.00-200 y=84.04x+349.78 0.9991 0.50 1.00 1.66 ± 0.03
Supplementary Table 2 Plasmatic and urinary biomarkers for the OACS model
Samples Ion (m/z) ESI mode Biomarker identification Molecular formula
Plasma
120.0818 + Phenylalanine C9H11NO2
232.1544 + Butyrylcarnitine C11H21NO4
188.0718 + Tryptophan C11H12N2O2
355.2632 + CA C24H40O5
496.3409 + LPC 16:0 C24H50NO7P
522.3562 + LPC 18:1 C26H52NO7P
524.3725 + LPC 18:0 C26H54NO7P
187.0073 - Cresol sulfate C7H8O4S
Urine
188.99 - Citrate C6H5O7
188.99 - Isocitrate C6H5O7
151.04 - 4-hydroxyphenylacetate C8H7O3
178..05 - Hippurate C9H9NO3
107.05 - 4-methyl-phenol C7H8O
76.04 + Trimethylamine-N-oxide C3H9NO
Supplementary Table 3 Main pharmacokinetics parameters of the eleven ginsenosides in the four group rats after i.g. administration of ginsenoside extracts (n=6)
Parameter Re Rg1 Rb1 20(S)-Rg2 Rc Rb2 Rd 20(S)-Rg3 CK
Cmax (ng/mL)
Blank 3.71 ± 0.82 12.33 ± 12.05 5.66 ± 1.37 3.33 ± 3.12 a
2.20 ± 0.35 b
15.54 ± 2.78 17.52 ±3.76 18.52 ± 5.31 5.87 ± 2.87
5.77 ± 2.56
1.90 ± 0.99
Model 2.22 ± 1.08 3.66 ±1.37 5.68 ±3.38 2.29 ± 1.27
2.73 ± 1.12
16.67 ± 7.27 17.44 ± 6.90 11.67 ± 4.46 2.44 ± 0.96*
2.75 ± 0.87*
3.21 ± 2.97
OF 1.74 ± 0.99 6.03 ±6.30 5.48 ± 1.60 2.06 ± 0.59
2.44 ± 0.67
18.03 ± 4.22 18.54 ± 8.25 40.23 ± 76.03 4.71 ± 1.83+
3.20 ± 1.07
1.72 ± 0.64
GP 2.01 ± 2.00 7.11 ±2.30+ 7.24 ± 3.38 3.62 ± 2.84
3.30 ± 0.61
24.94 ± 8.34 24.09 ± 10.00 20.54 ± 5.13+ 6.36 ± 2.51+
10.16 ± 4.56+
2.49 ± 1.37
Tmax (h)
Blank 0.39 ± 0.26 0.42 ± 0.29 6.50 ± 3.56 2.17 ± 1.47
8.67 ± 1.63
6.17 ± 4.12 7.50 ± 3.56 9.00 ± 1.67 2.20 ± 1.64
8.33 ± 0.82
10.33 ± 1.97
Model 0.39 ± 0.26 0.84 ± 0.19 8.00 ± 2.45 1.83 ± 1.17
8.33 ± 2.34
6.67 ± 2.42 5.67 ± 2.94 6.80 ± 2.68 2.33 ± 1.37
8.33 ± 2.34
8.67 ± 2.42
OF 0.28 ± 0.09 0.28 ± 0.09 2.93 ± 2.89 1.53 ± 0.65
7.60 ± 3.58
4.40 ± 3.29 4.40 ± 2.61 4.40 ± 2.61 1.53 ± 0.65
7.20 ± 3.35
8.80 ± 2.28
GP 0.34 ± 0.24 0.25 ± 0.09 2.00 ± 0.00 1.50 ± 0.58 5.00 ± 3.46 3.00 ± 1.16 5.00 ± 2.58 1.75 ± 0.96 9.50 ± 3.00
7.00 ± 1.16 7.00 ± 1.16
t 1/2 (h)
Blank 1.88 ± 0.69 4.24 ± 2.31 20.08 ± 8.23 36.75 ± 3.55 24.92 ± 15.37 31.91 ± 12.46 20.19 ± 6.60 24.03 ± 9.62 49.393 ± 15.64
Model 2.44 ± 1.42 3.25 ± 2.48 18.35 ± 12.64 38.65 ± 4.19 15.76 ± 4.98 19.41 ± 10.48 10.76 ± 6.71 26.18 ± 6.80 45.75 ± 24.55
OF 3.69 ± 1.77 1.95 ± 0.89 15.81 ± 5.52 34.74 ± 2.69 19.54 ± 6.15 20.10 ± 8.62 18.26 ± 11.43 16.35 ± 10.62 20.90 ± 12.86
GP 2.51 ± 1.34 1.60 ± 0.56 12.99 ± 4.68 35.73 ± 1.86 17.70 ± 8.43 21.45 ± 3.91 26.63 ± 3.78 30.37 ± 10.86 52.63 ± 14.447
AUC (ng·min/mL)
Blank 5.33 ± 1.35 29.79 ±32.46 161.22 ± 48.58 93.36 ± 17.58 447.35 ± 119.98 621.22 ± 114.03 374.62 ± 133.91 126.66 ± 31.23 63.43 ± 11.73
Model 3.04 ±1.37 7.36 ± 1.84 134.45 ± 92.45 65.66 ± 16.00* 414.01 ± 231.79 464.84 ± 239.85 195.27 ± 126.23* 67.47 ± 30.60** 57.68 ± 25.39
OF 1.89 ± 0.41 7.22 ± 2.20 133.56 ± 50.60 91.70 ± 11.65+ 468.52 ± 130.70 497.09 ± 181.79 251.32 ± 249.90 81.52 ± 21.74 57.45 ± 12.78
GP 2.19 ± 0.89 10.54 ± 4.31 172.70 ± 97.54 100.36 ± 23.21+ 542.29 ± 280.07 659.09 ± 236.66 431.74 ± 143.71+ 152.32 ± 27.23++ 75.99 ± 24.84 a: The value of the first peak; b: The value of the second peak; *: p<0.05, **: p<0.01, compared with Blank; +: p<0.05, ++: p<0.01, compared with Model.
Supplementary Table 4 Ginsenosides and relevant metabolites identified in the 24 h feces of the four group rats
Name tR (time) Molecular formula Characteristic ions
Rg1 7.12 C42H72O14 845.4915 [M-H+HCOOH]- (845.4899, 1.9); 799.4845 [M-H]- (799.4844, 0.1); 637.4335
[M-H-(Glc-H2O)]- (637.4316, 3.0); 475.3773 [M-H-2(Glc-H2O)]- (475.3787, -2.9)
Re 7.13 C48H82O18 991.5497 [M-H+HCOOH]-(991.5478, 1.9); 945.5436 [M-H]- (945.5423, 1.4); 799.4835
[M-H-(Rha-H2O)]- (799.4844, -1.1); 783.4887 [M-H-(Glc-H2O)]- (783.4895, -1.0); 637.4308
[M-H-(Rha-H2O)-(Glc-H2O)]- (637.4316, -1.3); 475.3790 [M-H-(Rha-H2O)-2(Glc-H2O)]- (475.3787, 0.6)
Rf 10.25 C42H72O14 845.4920 [M-H+HCOOH]- (845.4899, 2.5); 799.4851 [M-H]- (799.4844, 0.9); 637.4315
[M-H-(Glc-H2O)]- (637.4316, -0.2); 475.3781 [M-H-2(Glc-H2O)]- (475.3787, -1.3)
20(S)-Rg2 11.34 C42H72O13 829.4954 [M-H+HCOOH]- (829.4949, 0.6); 783.4888 [M-H]- (783.4895, -0.9); 637.4324
[M-H-(Rha-H2O)]- (637.4316, 1.3); 475.3786 [M-H-(Rha-H2O)-(Glc-H2O)]- (475.3787, -0.2)
Rb1 11.46 C54H92O23 1153.6012[M-H+HCOOH]- (1153.6006, 0.5); 1107.5961[M-H]- (1107.5951, 0.9)
20(S)-Rh1 11.48 C36H62O9 683.4373 [M-H+HCOOH]- (683.4370, 0.4); 637.4316 [M-H]- (637.4316, 0.0); 475.3793
[M-H-(Glc-H2O)]- (475.3787, 1.3)
20(R)-Rg2 11.53 C42H72O13 829.4957 [M-H+HCOOH]- (829.4949, 1.0); 783.4899 [M-H]- (783.4895, 0.5); 637.4315
[M-H-(Rha-H2O)]- (637.4316, -0.2); 475.3795 [M-H-(Rha-H2O)-(Glc-H2O)]- (475.3787, 1.7)
20(R)-Rh1 11.86 C36H62O9 683.4370 [M-H+HCOOH]- (683.4370, 0.0); 637.4299 [M-H]- (637.4316, -2.7); 475.3783
[M-H-(Glc-H2O)]- (475.3787, -0.8)
Rc 11.90 C53H90O22 1123.5905 [M-H+HCOOH]- (1123.5900, 0.4); 1077.5853 [M-H]- (1077.5845, 0.7); 945.5422
[M-H-(Ara(f)-H2O)]- (945.5423, -0.1)
Ro 11.93 C48H76O19 955.4907 [M-H]- (955.4903, 0.4); 793.4382 [M-H-(Glc-H2O)]- (793.4374, 1.0)
Rb2 12.33 C53H90O22 1123.5901 [M-H+HCOOH]- (1123.5900, 0.1); 1077.5854 [M-H]- (1077.5845, 0.8); 945.5423
[M-H-(Ara(f)-H2O)]- (945.5423, 0.0)
Rb3 12.46 C53H90O22 1123.5894 [M-H+HCOOH]- (1123.5900, -0.5); 1077.5845 [M-H]- (1077.5845, 0.0); 945.5416
[M-H-(Ara(f)-H2O)]- (945.5423, -0.7)
F1 12.88 C36H62O9 683.4368 [M-H+HCOOH]- (683.4370, -0.3); 637.4319 [M-H]- (637.4316, 0.5); 475.3788
[M-H-(Glc-H2O)]- (437.3787, 0.2)
Rd 13.30 C48H82O18 991.5471 [M-H+HCOOH]-(991.5478, -0.7); 945.5400 [M-H]- (945.5423, -2.4); 799.4835
[M-H-(Rha-H2O)]- (799.4844, -1.1); 783.4889 [M-H-(Glc-H2O)]- (783.4895, -0.8); 621.4360
[M-H-2(Glc-H2O)]- (621.4366, -1.0)
F2 16.65 C42H72O13 829.4960 [M-H+HCOOH]- (829.4949, 1.3); 783.4891 [M-H]- (783.4895, -0.5); 621.4369
[M-H-(Glc-H2O)]- (621.4366, 0.5); 459.3834 [M-H-2(Glc-H2O)]- (459.3838, -0.9)
20(S)-Rg3 17.68 C42H72O13 829.4953 [M-H+HCOOH]- (829.4949, 0.5); 783.4897 [M-H]- (783.4895, 0.3); 621.4363
[M-H-(Glc-H2O)]- (621.4366, -0.5); 459.3831 [M-H-2(Glc-H2O)]- (459.3838, -1.5)
20(S)-PPT 18.67 C30H52O4 521.3840 [M-H+HCOOH]- (521.3842, -0.4)
CK 21.92 C36H62O8 667.4415 [M-H+HCOOH]- (667.4421, -0.9); 621.4352 [M-H]- (621.4366, -2.3); 459.3826
[M-H-(Glc-H2O)]- (459.3838, -2.6)
20(S)-Rh2 22.42 C36H62O8 667.4427 [M-H+HCOOH]- (667.4421, 0.9); 621.4382 [M-H]- (621.4366, 2.6)
20(R)-Rh2 22.76 C36H62O8 667.4425 [M-H+HCOOH]- (667.4421, 0.6); 621.4351 [M-H]- (621.4366, -2.4)
Rg6 15.64 C42H70O12 811.4838 [M-H+HCOOH]- (811.4844, -0.7); 765.4789 [M-H]- (765.4789, 0.0)
F4 15.95 C42H70O12 811.4838 [M-H+HCOOH]- (811.4844, -0.7); 765.4774 [M-H]- (765.4789, -2.0)
Rk3 16.24 C36H60O8 665.4264 [M-H+HCOOH]- (665,4265, -0.2)
Rh4 16.67 C36H60O8 665.4265 [M-H+HCOOH]- (665,4265, 0.0)
20(R)-Rg3 18.25 C42H72O13 829.4944[M-H+HCOOH]- (829.4949, -0.6); 783.4891[M-H]- (783.4895, -0.5);
621.4373[M-H-(Glc-H2O)]- (621.4366, 1.1); 459.3837 [M-H-2(Glc-H2O)]- (459.3838, -0.2)
20(R)-PPT 18.70 C30H52O4 521.3850 [M-H+HCOOH]- (521.3842, 1.5)
Rk1 21.27 C42H70O12 811.4848 [M-H+HCOOH]- (811.4844, 0.5); 765.4783 [M-H]- (765.4789, -0.8)
Rg5 21.67 C42H70O12 811.4853 [M-H+HCOOH]- (811.4844, 1.1); 765.4780 [M-H]- (765.4789, -1.2)
Supplementary Table 5 Contents of the eleven ginsenosides in the 24 h feces of the four group rats (ng/g, n=6)
Analyte Blank Model OF GPs
Re 6093.60 ± 6348.53 121491.70 ± 104632.70* 18243.78 ± 15545.76+ 23477.91 ± 15355.90+
Rg1 218534.90 ± 315766.60 58256.36 ± 60478.57 322962.40 ± 582047.00 67117.65 ± 62744.95
Rf 647.087 ± 470.41 442.13 ± 314.67 783.65 ± 923.21 514.78 ± 226.39
Rb1 3.71 ± 2.43 2.49 ± 1.84 84.74 ± 156.73 9.19 ± 20.30
20(S)-Rg2 16302.83 ± 10154.32 72229.66 ± 60461.32* 5444.08 ± 5507.11+ 13217.92 ± 10363.85+
Rc 45.13 ± 28.75 72.95 ± 35.02* 16.05 ± 8.66++ 47.65 ± 13.01+
Rb2 54.89 ± 39.70 99.62 ± 311.62 1346.50 ± 2637.37 60.93 ± 72.10
Rd 1410.65 ± 668.07 645.57 ± 484.50** 218.59 ± 165.28++ 299.44 ± 124.13+
F2 3063.95 ± 3334.22 1976.36 ± 1090.99 34987.77 ± 55533.93 2392.80 ± 1792.33
20(S)-Rg3 236301.80 ± 64669.82 201318.50 ± 137314.20 376070.00 ± 426707.80 95815.06 ± 55398.23+
CK 714344.80 ± 175067.20 789820.10 ± 190996.60 415378.90 ± 38426.26++ 470279.30 ± 173142.50++ *: p<0.05, **: p<0.01, compared with Blank; +: p<0.05, ++: p<0.01, compared with Model.
Supplementary Table 6 Analytical conditions for quantitative determination of the eleven ginsenosides by HPLC-TQ-MS
Analyte tR (min) Channel MRM Cone voltage (V) Collision voltage (eV)
Re 1.92 Ⅰ
m/z 969 → m/z 203 75 55
Rg1 1.94 m/z 823 → m/z 203 65 40
Digoxin 5.39 Ⅱ
m/z 803 → m/z 283 70 50
Rf 6.08 m/z 823 → m/z 365 70 50
Rb1 6.73
Ⅲ
m/z 1131 → m/z 365 80 60
20(S)-Rg2 7.07 m/z 807 → m/z 349 75 45
Rc 7.08 m/z 1101 → m/z 335 65 60
Rb2 7.41 m/z 1101 → m/z 335 65 60
Rd 8.17 Ⅳ m/z 969 → m/z 203 70 50
F2 10.55
Ⅴ
m/z 807 → m/z 203 70 45
20(S)-Rg3 11.57 m/z 807 → m/z 365 80 50
CK 14.29 m/z 645 → m/z 203 50 35
Supplementary Table 7 Linearity and sensitivity for the eleven ginsenosides in rat plasma (n=3)
Analyte Linear range (ng/mL) Equationa R2 LOD (ng/mL) LOQ (ng/mL)
Re 0.25-100 y=0.0069x+0.0028 0.9998 0.10 0.25
Rg1 0.25-100 y=0.0138x+0.0052 0.9992 0.10 0.25
Rf 0.25-100 y=0.0040x+0.0008 0.9994 0.10 0.25
Rb1 0.25-100 y=0.6417x-0.0209 0.9997 0.10 0.25
20(S)-Rg2 0.25-100 y=0.0021x+0.0011 0.9996 0.10 0.25
Rc 0.25-100 y=0.0347x+0.0042 0.9994 0.10 0.25
Rb2 0.25-100 y=0.0431x+0.0095 0.9995 0.10 0.25
Rd 0.25-100 y=0.0145x-0.0014 0.9993 0.10 0.25
F2 0.25-100 y=0.0294x+0.0099 0.9996 0.10 0.25
20(S)-Rg3 0.25-100 y=0.0079x-0.0017 0.9990 0.10 0.25
CK 0.25-100 y=0.0089x+0.0008 0.9991 0.10 0.25 a: mean of three standard curves
Supplementary Table 8 Matrix effects, intra-, inter-day precisions and accuracies for the eleven ginsenosides in rat plasma (n=3)
Analyte QC concentration
(ng/mL)
Matrix effect
Intra-day Inter-day
Mean (%) RSD (%) Precision (RSD %) Accuracy (RE %) Precision (RSD %) Accuracy (RE %)
Re 0.50 87.34 9.01 5.62 -9.56 11.66 +5.55
2.50 95.97 8.10 5.33 -5.87 5.33 -9.85
10.00 97.62 4.83 8.75 +5.10 7.56 -0.53
80.00 85.18 1.46 5.62 -5.67 7.22 -11.54
Rg1 0.50 88.05 6.10 5.88 -3.61 7.11 -8.85
2.50 94.22 7.75 10.49 +7.21 4.59 -9.40
10.00 87.93 4.00 4.46 -14.20 6.74 -11.20
80.00 98.56 1.64 6.13 +7.48 9.32 +13.42
Rf 0.50 89.49 3.94 12.91 -9.64 12.57 -8.75
2.50 97.84 2.94 1.59 -12.83 13.72 -0.12
10.00 94.69 1.49 6.81 -8.10 9.13 -14.09
80.00 90.78 0.61 1.80 -13.85 9.37 -14.97
Rb1 0.50 98.90 1.03 2.98 -1.84 5.35 -1.28
2.50 98.93 3.38 9.63 -1.54 10.89 -4.69
10.00 96.88 2.97 5.33 -14.28 6.73 -14.62
80.00 87.45 0.58 1.12 -13.61 5.41 -9.93
20(S)-Rg2 0.50 89.33 6.72 13.22 -10.47 12.55 -9.98
2.50 87.49 0.97 10.88 -3.46 10.88 -2.75
10.00 91.04 8.50 5.36 -4.06 3.94 -10.06
80.00 95.23 5.46 0.57 +0.47 6.90 +3.53
Rc 0.50 91.73 1.48 5.67 -4.22 5.27 -4.33
2.50 94.69 3.90 13.45 -9.90 5.96 -0.03
10.00 96.14 5.90 5.96 -9.79 5.91 -3.36
80.00 98.85 1.02 8.98 -4.81 12.83 -8.04
Rb2 0.50 96.49 5.47 13.42 -4.07 12.39 -0.05
2.50 93.56 3.09 14.01 -12.32 9.86 -5.26
10.00 98.24 2.95 3.48 -14.56 6.37 -12.54
80.00 85.36 0.57 1.37 -13.17 3.49 -14.65
Rd 0.50 92.05 1.23 7.17 -7.86 4.98 -7.02
2.50 94.70 8.64 9.48 -14.18 12.00 -14.60
10.00 91.00 2.08 9.51 -0.61 7.80 +1.28
80.00 89.35 4.51 1.04 -12.52 1.85 -10.95
F2 0.50 88.89 3.82 4.43 -8.87 4.82 -12.82
2.50 98.77 7.73 5.07 -13.72 3.59 -9.92
10.00 97.80 6.69 0.12 -11.71 7.55 -10.17
80.00 85.92 0.10 3.59 -8.23 3.59 -8.33
20(S)-Rg3 0.50 85.26 2.06 3.68 -7.91 2.84 -6.53
2.50 90.94 5.20 11.37 -9.86 12.95 -12.85
10.00 95.44 1.72 4.07 -9.83 5.98 -8.09
80.00 87.52 1.94 0.90 -10.39 0.90 -11.19
CK 0.50 86.14 2.88 3.28 +0.37 2.46 -6.08
2.50 90.79 8.67 3.88 +4.89 4.29 +3.56
10.00 90.71 9.69 3.89 +7.89 3.39 +0.23
80.00 96.34 4.39 11.87 -15.85 7.79 -9.99
Supplementary Table 9 Recovery and stability of the eleven ginsenosides in rats plasma (n=3)
Analyte
QC
concentration
(ng/mL)
Recovery
Stability (RSD, %)
Mean (%) RSD (%) Freeze/thaw stability
(-20°C to 25°C)
Autosampler stability
(10°C)
Short-term stability
(25°C)
Long-term stability
(-20°C)
Re 0.50 95.95 8.08 2.53 11.35 14.70 6.93
2.50 89.43 8.10 9.92 11.30 5.33 8.43
10.00 94.03 4.83 3.43 6.94 8.75 4.36
80.00 92.40 6.20 1.74 3.54 5.62 4.24
Rg1 0.50 89.56 8.97 8.87 6.55 9.73 13.85
2.50 90.16 8.94 9.68 2.72 5.15 8.94
10.00 90.31 5.94 8.88 1.92 4.46 8.10
80.00 99.10 1.64 8.72 7.84 6.13 1.49
Rf 0.50 86.31 13.70 12.92 12.66 14.34 14.03
2.50 87.11 3.08 4.64 8.97 1.59 8.26
10.00 91.29 7.55 8.80 5.84 6.81 4.22
80.00 85.81 0.55 6.45 0.83 1.80 3.77
Rb1 0.50 93.96 0.92 3.86 5.81 1.87 0.59
2.50 91.74 3.58 7.93 10.18 0.43 8.52
10.00 89.16 6.34 3.95 4.03 5.33 4.46
80.00 86.48 0.85 5.77 1.12 3.71 1.22
20(S)-Rg2 0.50 88.25 5.98 12.53 12.94 14.50 7.44
2.50 83.55 0.97 3.14 9.26 8.35 4.70
10.00 89.29 9.55 2.41 1.80 5.36 3.06
80.00 97.64 3.62 1.02 8.06 0.57 1.22
Rc 0.50 90.28 2.78 6.39 5.57 4.49 5.19
2.50 99.35 7.31 8.72 5.96 5.01 9.43
10.00 97.68 8.16 5.61 3.51 5.96 7.20
80.00 95.62 9.89 9.04 9.99 8.98 6.78
Rb2 0.50 88.15 5.68 3.70 14.22 5.74 5.03
2.50 98.25 9.47 2.78 7.10 4.56 7.07
10.00 88.44 4.57 6.77 0.35 3.48 1.12
80.00 86.26 0.54 2.61 0.34 1.37 4.33
Rd 0.50 86.82 2.47 2.46 5.73 0.98 2.82
2.50 92.82 12.73 9.48 10.72 9.48 5.81
10.00 92.07 2.08 7.68 0.49 9.51 6.03
80.00 88.11 4.13 1.53 2.26 1.04 1.22
F2 0.50 85.78 4.58 6.04 5.54 1.61 2.99
2.50 90.32 6.92 3.91 0.23 9.54 5.58
10.00 95.69 5.98 9.72 2.96 0.12 5.65
80.00 91.39 3.90 3.01 2.02 3.59 6.41
20(S)-Rg3 0.50 89.27 1.84 3.14 3.33 1.26 1.38
2.50 86.61 1.25 9.56 0.62 7.99 3.76
10.00 91.41 4.71 5.49 5.50 4.07 8.42
80.00 88.39 1.76 5.26 4.79 0.90 2.43
CK 0.50 93.27 3.37 2.24 2.01 0.68 3.13
2.50 98.12 5.26 3.49 6.70 3.88 2.57
10.00 96.35 8.37 10.62 3.39 9.06 6.65
80.00 85.99 3.94 7.84 3.28 6.42 6.15
Supplementary Figures
Gut microbiota-involved mechanisms in enhancing systemic exposure of ginsenosides by coexisting
polysaccharides in ginseng decoction
Shan-Shan Zhou1, 2, #, Jun Xu3, #, He Zhu1, Jie Wu2, Jin-Di Xu1, Ru Yan4, Xiu-Yang Li1, 2, Huan-Huan Liu1, 2, Su-Min Duan2,
Zhuo Wang2, Hu-Biao Chen3, Hong Shen2, *, Song-Lin Li1, 2, *
Supplementary figure legends
Supplementary Figure 1 Molecular weight distribution (a) and compositional monosaccharide analysis (b: mixed standards; c: sample) of ginseng polysaccharides
Supplementary Figure 2 Chemical structures of ginsenosides and digoxin (the internal standard)
Supplementary Figure 3 OPLS-DA (a: score plot; b: loading plot; c: S-plot) results based on the plasma metabolic profiling of rats in negative (1) and positive (2) ion modes (♦: Blank; +: Model; n=6)
Supplementary Figure 4 PCA (a: score plot) and OPLS-DA (b: loading plot; c: S-plot) results based on the urine metabolic profiling of rats in negative (1) and positive (2) ion modes (♦: Blank; +: Model; n=6)
Supplementary Figure 5 Comparison of potential biomarkers in the plasma (a) and urine (b) between the four group rats (n=6);*: p<0.05, **: p<0.01, compared with Blank; +: p<0.05, ++: p<0.01, compared with Model
Supplementary Figure 6 Species accumulation curves (a) and Shannon-Wiener curves (b) of the samples
Supplementary Figure 7 Chemical profiling of 24 h feces in the four group rats after i.g. administration of ginsenoside extracts by UPLC-QTOF-MS analysis
b
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Supplementary Figure 1
Supplementary Figure 2
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Supplementary Figure 6
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Supplementary Figure 7